Project Summary The main objective of Project 2 is to identify and characterize cortical neurons from layer V and subtypes of astrocytes derived from human induced pluripotent stem cells (iPSCs) and measure their contribution to Autism Spectrum Disorders (ASD). Pyramidal neurons from cortical layer V are linked to primate social behavior and human-related conditions, such as ASD and Williams syndrome (WS). Glia cells have historically played a secondary role in neuroscience but recent data indicating a major contribution of astrocytes to several human neurological disorders has challenged this view. Thus, we propose to apply the design of experiments (DOE), a powerful statistical tool, to improve the differentiation protocols for cortical neurons layer V and astrocytes derived from iPSCs. These will be applied to engineered lines from Project 1 for molecular and cellular assays in the Projects 3, 4 and 5.